Beta-lactam inhibitors of CoA-IT

ABSTRACT

This invention relates to methods for treating diseases or disorders mediated by lipid inflammatory mediators, arachidonic acid, its metabolites and/or platelet activating factor (PAF), which method comprises administration to a mammal in need thereof of an effective Coenzyme A independent transacylase (CoA-IT) inhibiting amount of a triphenylmethylazetidinone. This invention also relates to a method of treating or reducing inflammation in a mammal in need thereof, which comprises administering to said mammal an effective amount of a triphenylmethylazetidinone.

This application is a 371 of PCT/US 98/09483, filed May 8, 1998, andclaims priority to provisional application 60/046,080, filed May 9,1997.

FIELD OF THE INVENTION

This invention relates to novel compounds, pharmaceutical compositionsthereof, and their use as anti-inflammatory agents in mammals.

BACKGROUND OF THE INVENTION

Coenzyme A-independent transacylase (CoA-IT) is an enzyme responsiblefor the movement of arachidonate between phospholipid molecular speciesof inflammatory cells. CoA-IT removes arachidonate from the sn-2position of 1-acyl-containing phospholipids, such as1-acyl-2-arachidonoyl-sn-glycero-3-phosphocholine(1-acyl-2-arachidonoyl-GPC). It then transfers that arachidonate to asuitable lyso-phospholipid acceptor, such as 1-alkyl-2-lyso-GPC and1-alkenyl-2-lyso-sn-glycero-3-phospho-ethanolamine (Sugiura et al., J.Biol. Chem. 262: 1199-1205 (1987); Kramer and Deykin, Biol. Chem. 258:13806-13811 (1983); Chilton et al., J. Biol. Chem. 258: 7268-7271(1983)). This activity is selective for 20 carbon fatty acyl groups andis the mechanism by which inflammatory cells move arachidonate intospecific phospholipid pools prior to its release (Winkler and Chilton,Drug News Perspec. 6: 133-138 (1993); Snyder et al., J. Lipid Mediat.10: 25-31 (1994)).

Further, a method which antagonises the production of free arachidonicacid, its metabolites or PAF will have clinical utility in the treatmentof a variety of allergic, inflammatory and hypersecretory conditionssuch as asthma, arthritis, rhinitis, bronchitis and urticaria, as wellas reperfusion injury and other disease involving lipid mediators ofinflammation. Many published patent applications or issued U.S. patentsexist which describe various compounds having utility as PAF oreicosanoid antagonists. Such patents include U.S. Pat. Nos. 4,788,205,4,801,598, 4,981,860, 4,992,455, 4,983,592, 5,011,847, 5,019,581 and5,002,941.

Accordingly, as CoA-IT is involved in arachidonic acid and phospholipidmetabolism, inhibition of such an enzyme would be useful for thetreatment of inflammatory, allergic and hypersecretory conditions ordisease states caused thereby. Therefore, a method by which CoA-IT isinhibited will consequently and preferentially decrease the arachidonatecontent of 1-alkyl- and 1-alkenyl-linked phospholipids and willtherefore decrease the production of pro-inflammatory mediators such asfree arachidonic acid, prostaglandins, leukotriene and PAF during aninflammatory response.

There remains a need for treatment, in this field, for compounds whichare CoA-IT inhibitors, i.e. compounds which are capable of inhibiting,or interfering with this enzyme and thereby decrease production of thepro-inflammatory mediators.

SUMMARY OF THE INVENTION

This invention also relates to a method of treating or reducinginflammation in a mammal in need thereof, which comprises administeringto said mammal an effective amount of a compound or composition ofFormula (I).

This invention also relates to a method of treating disease or disordersmediated by lipid inflammatory mediators, free arachidonic acid, itsmetabolites and/or PAF by administering to a patient in need thereof, aneffective amount of a compound of Formula (I).

This invention also relates to a method of treating disease or disordersmediated by Coenzyme A independent transacylase (CoA-IT) byadministering to a patient in need thereof, an effective amount of acompound or composition of Formula (I).

This invention relates to the novel compounds of Formula (Ia) andpharmaceutically acceptable salts thereof. The present invention alsoprovides for a pharmaceutical composition comprising a pharmaceuticalacceptable carrier or diluent and a compound of Formula (Ia), orpharmaceutically acceptable salt thereof.

One aspect of the present invention are the compounds represented by astructure having the formula:

wherein

Y is NH;

X is O or S(O)m;

m is 0 or an integer having a value of 1, or 2;

R₃ is optionally substituted triphenylmethyl;

R₄ is optionally substituted C₁₋₁₀ alkyl, (CR₁₀R₂₀)_(n)C(R₁₀)=C(R₇)₂, or(CR₁₀R₂₀)n—C≡C—R₅;

n is an integer having a value of 1 to 4;

R₁₀ and R₂₀ are independently hydrogen or C₁₋₄ alkyl;

R₅ is hydrogen, alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,C(O)₂R₆, or C(O)R₆ wherein the alkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl moieties may be optionally substituted;

R₆ is C₁₋₁₀ alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,heterocyclic, or heterocyclicalkyl, all of which may be optionallysubstituted:

R₇ is independently hydrogen. C₁₋₁₀ alkyl, aryl, arylalkyl, heteroaryl.heteroarylalkyl, heterocyclic, or heterocyclicalkyl, all of which may beoptionally substituted;

or a pharmaceutically acceptable salt thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 demonstrates the time dependent inhibition of CoA-IT by(3S,4R)-4-(isobutenyloxy)-3-(triphenylmethylamino)azetidin-2-one.

FIG. 2 demonstrates inhibition of PAF production in neutrophils by(3S,4R)-4-(isobutenyloxy)-3-(triphenylmethylamino)azetidin-2-one.

FIG. 3 demonstrates inhibition of LTC₄ production in monocytes by(3S,4R)-4-(isobutenyloxy)-3-(triphenylmethylamino)azetidin-2-one.

FIG. 4 demonstrates inhibition of PGE₂ production in monocytes by(3S,4R)-4-(isobutenyloxy)-3-(triphenylmethylamino)azetidin-2-one.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is directed to a novel method of treatinginflammatory disease in a mammal in need thereof by administering tosaid mammal an effective amount of a compound according to Formula (I).The compounds of Formula (I) selectively inhibit the CoA-IT enzyme. Thiswill result in the treatment of inflammatory occurrences in mammals.Inflammatory states in mammals may include, but are not limited to,allergic and asthmatic manifestations, dermatological diseases,inflammatory diseases, collagen diseases, reperfusion injury and stroke.Treatment of both acute and chronic diseases are possible. Preferreddiseases for treatment are arthritis, asthma, allergic rhinitis,inflammatory bowel disease (IBD), psoriasis, reperfusion injury andstroke. For the purposes herein, the compounds of Formula (I) arepreferential and selective inhibitors of CoA-IT.

Suitably, in compounds of Formula (I), X is O or S(O)m; and m is 0 or aninteger having a value of 1, or 2. Preferably m is 0 or 2.

Suitably, in compounds of Formula (I), R₃ is optionally substitutedtriphenylmethyl group. The phenyl rings may be independently substitutedone to three times by halogen, such as fluorine, chlorine, bromine oriodine; hydroxy; hydroxy substituted C₁₋₁₀ alkyl; C₁₋₁₀ alkoxy, such asmethoxy or ethoxy; halosubstituted C₁₋₁₀ alkoxy; S(O)m alkyl, such asmethyl thio, methylsulfinyl or methyl sulfonyl; S(O)m aryl; amino, mono& di- C₁₋₁₀ alkyl substituted amino; C₁₋₁₀ alkyl; halosubstituted C₁₋₁₀alkyl, such as CF₃; CHO, C(O)C₁₋₁₀ alkyl, C(O) aryl, C(O)₂R₈, wherein R₈is C₁₋₁₀ alkyl, aryl, or arylalkyl; C(O)NR₉R₁₁; cyano; S(O)₂ NR₀R₁₁;N(R₁₀)C(O)R₆; N(R₁₀)C(O) NR₉R₁₁; N(R₁₀)C(O)OR₆; or N(R₁₀)S(O)₂R₆.

Suitably, R₉ and R₁₁ are independently hydrogen, C₁₋₁₀ alkyl, aryl,arylalkyl.

Suitably, R₄ is optionally substituted C₁₋₁₀ alkyl(CR₁₀R₂₀)_(n)C(R₁₀)=C(R₇)₂, or (CR₁₀R₂₀)n—C≡C—R₅; wherein n is aninteger having a value of 1 to 4. Preferably n is 1.

When R₄ is an optionally substituted C₁₋₁₀ alkyl, the alkyl moiety maybe straight or branched, and may be substituted one or more times,independently by halogen, such as fluorine; hydroxy; C₁₋₁₀ alkoxy; S(O)malkyl, wherein m is 0, 1 or 2; amino, mono & di-substituted amino, suchas NR₉R₁₁ group; wherein R₉ and R₁₁ are as described above, or R₉ andR₁₁ together with the nitrogen to which they are attached cyclize toform a 5 to 7 membered ring which optionally includes an additionalheteroatom selected from O/N/S; —O(CR₁₀R₂₀)_(s)O— wherein s is aninteger having a value of 2 to 4 and both oxygens are attached to thesame carbon in R₄; —S(CR₁₀R₂₀)_(s)S— wherein s is as previously definedand both sulfurs are attached to the same carbon in R4; cycloalkyl, or acycloalkyl alkyl group; a halosubstituted C₁₋₄ alkyl, such as CF₃; anoptionally substituted aryl, such as phenyl, or an optionallysubstituted arylalkyl, such as benzyl or phenethyl, heteroaryl, orheteroarylalkyl, wherein these aryl or heteoraryl moieties may also besubstituted one to two times by halogen; hydroxy; hydroxy substitutedalkyl; C₁₋₁₀ alkoxy; S(O)_(m) alkyl; amino, mono & di-C₁₋₄ alkylsubstituted amino, such as in the NR₉R₁₁ group (wherein R₉ and R₁₁ areas defined above); C₁₋₁₀ alkyl, or CF₃.

Preferably, R₄ is a C₁₋₄ alkyl, such as isobutyl, or an alkenyl, such asisobutenyl.

Suitably, R₁₀ and R₂₀ are independently hydrogen or C₁₋₄ alkyl.

Suitably, R₅ is hydrogen, C₁₋₁₀ alkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, C(O)₂R₆, C(O)R₆. Preferably R₅ is hydrogen, C(O)₂R₆, ora heteroaryl ring, and preferably R₆ therein is a C 1-4 alkyl, such asmethyl. If R₅ is a heteroaryl ring, it is preferably a 2-, 3,- or4-pyridyl. The alkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkylmay be optionally substituted as herein defined.

Suitably, R₆ is C₁₋₁₀ alkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, heterocyclic, or heterocyclicalkyl. The aryl,arylalkyl, heteroaryl, heteroarylalkyl, heterocyclic, andheterocyclicalkyl moieties may be optionally substituted as hereindefined.

Suitably, R₇ is independently hydrogen, C₁₋₁₀ alkyl, aryl, arylalkyl,heteroaryl, heteroarylalkyl, heterocyclic, or heterocyclicalkyl. Thealkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclic, andheterocyclicalkyl moieties may be optionally substituted as hereindefined.

Suitable pharmaceutically acceptable salts are well known to thoseskilled in the art and include basic salts of inorganic and organicacids, such as hydrochloric acid, hydrobromic acid, sulphuric acid,phosphoric acid, methane sulphonic acid, ethane sulphonic acid, aceticacid, malic acid, tartaric acid, citric acid, lactic acid, oxalic acid,succinic acid, fumaric acid, maleic acid, benzoic acid, salicylic acid,phenylacetic acid and mandelic acid

The following terms, as used herein, refer to:

“halo”—all halogens, that is chloro, fluoro, bromo and iodo;

“C₁₋₁₀ alkyl” or “alkyl”—both straight and branched chain radicals of 1to 10 carbon atoms, unless the chain length is otherwise limited,including, but not limited to, methyl, ethyl, n-propyl, iso-propyl,n-butyl, sec-butyl, iso-butyl, tert-butyl, and the like;

“cycloalkyl” is used herein to mean cyclic radicals, preferably of 3 to8 carbons, including but not limited to cyclopropyl, cyclopentyl,cyclohexyl, and the like;

“alkenyl” is used herein at all occurrences to mean straight or branchedchain radical of 2-10 carbon atoms, unless the chain length is limitedthereto, including, but not limited to ethenyl, 1-propenyl, 2-propenyl,2-methyl-1-propenyl, 1 -butenyl, 2-butenyl and the like;

“aryl”—phenyl and naphthyl;

“heteroaryl” (on its own or in any combination, such as “heteroaryloxy”,or “heteroaryl alkyl”)—a 5-10 membered aromatic ring system in which oneor more rings contain one or more heteroatoms selected from the groupconsisting of N, 0 or S, such as, but not limited, to pyrrole, pyrazole,furan, thiophene, quinoline, isoquinoline, quinazolinyl, pyridine,pyrimidine, oxazole, thiazole, thiadiazole, triazole, imidazole, orbenzimidazole;

“heterocyclic” (on its own or in any combination, such as“heterocyclylalkyl”)—a saturated or partially unsaturated 4-10 memberedring system in which one or more rings contain one or more heteroatomsselected from the group consisting of N, O, or S; such as, but notlimited to, pyrrolidine, piperidine, piperazine, morpholine,tetrahydropyran, or imidazolidine;

The term “aralkyl” or “heteroarylalkyl” or “heterocyclicalkyl” is usedherein to mean C₁₋₄ alkyl as defined above attached to an aryl,heteroaryl or heterocyclic moiety as also defined herein unlessotherwise indicate;

“sulfinyl”—the oxide S (O) of the corresponding sulfide, the term “thio”refers to the sulfide, and the term “sulfonyl” refers to the fullyoxidized S(O)₂ moiety;

“aroyl”—a C(O)Ar, wherein Ar is as phenyl, naphthyl, or aryl alkylderivative such as defined above, such group include but are notelimited to benzyl and phenethyl;.

“alkanoyl”—a C(O)C₁₋₁₀ alkyl wherein the alkyl is as defined above.

“optionally substituted” unless specifically defined for a particularsubstituent group, shall mean such groups as halogen, such as fluorine,chlorine, bromine or iodine; hydroxy; hydroxy substituted C₁₋₁₀ alkyl;C₁₋₁₀ alkoxy, such as methoxy or ethoxy; S(O)m alkyl, wherein m is 0, 1or 2, such as methyl thio, methylsulfinyl or methyl sulfonyl; amino,mono & di-substituted amino, such as in NR₉R₁₁ group; C₁₋₁₀ alkyl,cycloalkyl, or cycloalkyl alkyl group, such as methyl, ethyl, propyl,isopropyl, t-butyl, etc., cyclopropyl, or cyclopropyl methyl;halosubstituted C₁₋₁₀ alkyl, such as CF₃; an optionally substitutedaryl, such as phenyl, or an optionally substituted arylalkyl, such asbenzyl or phenethyl, wherein these aryl moieties may also be substitutedone to two times by halogen; hydroxy; hydroxy substituted alkyl; C₁₋₁₀alkoxy; S(O)_(m) alkyl; amino, mono & di-C₁₋₄ alkyl substituted amino,such as in the NR₉R₁₁ group; C₁₋₁₀ alkyl, or CF₃.

It is recognized that the compounds of the present invention may existas stereoisomers, regioisomers, or diastereoisomers. These compounds maycontain one or more asymmetric carbon atoms and may exist in racemic andoptically active forms. All of these compounds are included within thescope of the present invention.

Another aspect of the present invention are the novel compounds ofFormula (Ia) represented by the structure:

wherein

Y is NH;

X is O or S(O)m;

m is 0 or an integer having a value of 1, or 2;

R₃ is optionally substituted triphenylmethyl;

R₄ is optionally substituted C₁₋₁₀ alkyl, (CR₁₀R₂₀)_(n)C(R₁₀)=C(R₇)₂, or(CR₁₀R₂₀)n—C≡C—R₅;

n is an integer having a value of 1 to 4;

R₁₀ and R₂₀ are independently hydrogen or C₁₋₄ alkyl;

R₅ is hydrogen, alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,C(O)₂R₆, or C(O)R₆ wherein the alkyl, aryl, arylalkyl, heteroaryl andheteroarylalkyl moieties may be optionally substituted;

R₆ is C₁₋₁₀ alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,heterocyclic, or heterocyclicalkyl, wherein the alkyl, aryl, arylalkyl,heteroaryl, heteroarylalkyl, heterocyclic, or heterocyclicalkyl moietiesmay be optionally substituted;

R₇ is independently hydrogen, C₁₋₁₀ alkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, heterocyclic, or heterocyclicalkyl, wherein the alkyl,aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclic, andheterocyclicalkyl moieties may be optionally substituted;

excluding 4-methoxy-3-(triphenylmethylamino)azetidin-2-one,3-(triphenylmethylamino)-azetidin-2-one,4-(isobutenyloxy)-3-(triphenylmethylamino)azetidin-2-one,4-(methylsulfonyl)-3-(triphenylmethylamino)azetidin-2-one, and4-(prop-2-ynyloxy)-3-(triphenylmethylamino)azetidin-2-one;

or a pharmaceutically acceptable salt thereof.

For compounds of Formula (Ia), the variables Y, X, n, m, R₃, R₄, R₁₀,R₂₀, R₅, R₆, and R₇ are as defined above for compounds of Formula (I).

Specifically exemplified compounds of Formula (I) are:

(3RS,4RS)-4-(Isobutoxy)-3-(triphenylmethylamino)azetidin-2-one

(3R,4R)-4-(Isobutoxy)-3-(triphenylmethylamino)azetidin-2-one

(3S ,4S)-4-(Isobutoxy)-3-(triphenylmethylamino)azetidin-2-one

(3R,4R)-4-(Isobutylthio)-3-(triphenylmethylamino)azetidin-2-one

(3R,4R)-4-(Isobutylsulfonyl)-3-(triphenylmethylamino)azetidin-2-one

(3S,4R)-4-(Isobutoxy)-3-(triphenylmethylamino)azetidin-2-one

(3S,4R)-4-(Propoxy)-3-(triphenylmethylamino)azetidin-2-one

(3S ,4S)-4-(Propoxy)-3-(triphenylmethylamino)azetidin-2-one

(3S,4R)-4-(Benzyloxy)-3-(triphenylmethylamino)azetidin-2-one

(3S,4S)-4-(Benzyloxy)-3-(triphenylmethylamino)azetidin-2-one

(3S,4R)-4-Methoxy-3-(triphenylmethylamino)azetidin-2-one

(3S,4R)-4-(Isobutenyloxy)-3-(triphenylmethylamino)azetidin-2-one

(3S,4R)-4-Octyloxy-3-(triphenylmethylamino)azetidin-2-one

(3S ,4R)-4-Phenoxy-3-(triphenylmethylamino)azetidin-2-one

(3S,4S)-4-Phenoxy-3-(triphenylmethylamino)azetidin-2-one

(3S,4R)-3-[[(4-Iodophenyl)diphenylmethyl]amino]-4-(isobutoxy)azetidin-2-one

(3S,4S)-4-[3-(Methoxycarbonyl)propoxy]-3-(triphenylmethylamino)azetidin-2-one

(3S,4R)-4-[[2-(3-Pyridylmethyl)-1,3-dithian-2-yl]methoxy]-3-(triphenylmethylamino)-azetidin-2-one

(3S,4S)-4-(Prop-2-ynyloxy)-3-(triphenylmethylamino)azetidin-2-one

Methyl4-[(3S,4S)-2-oxo-3-(triphenylmethylamino)azetidin-4-yloxy]but-2-ynoate

Methyl4-[(3S,4R)-2-oxo-3-(triphenylmethylamino)azetidin-4-yloxy]but-2-ynoate

(3S,4R)-4-[(2(5H)Furanon-4-yl)methoxy]-3-(triphenylmethylamino)azetidin-2-one

(S)-3-(Triphenylmethylamino)azetidin-2-one

(RS)-3-(Triphenylmethylamino)azetidin-2-one

(3R,4R)-4-(Methylsulfonyl)-3-(triphenylmethylamino)azetidin-2-one

Compounds of formula (I) where 4-(X—R⁴) is 4-(O—R⁴) or 4-(S—R⁴) can beprepared according to Scheme I from 1, where R³ is defined as in formula(I) and Y is a suitable leaving group such as methylsulfonyl, acyloxy,or chloro. Compounds 1, where Y is methylsulfonyl, are obtained asdescribed in J. Chem. Soc. Perkin I, 447, 1976 whose disclosure isincorporated herein by reference. The displacement of group Y in 1 withHO—R⁴ or HS—R⁴ may be carried out with a suitable catalyst such as zincacetate in a suitable solvent such as toluene at a suitable temperaturesuch at 90° C. Alternatively, with phenols, the displacement may becarried out in the presence of a suitable base, such as aqueous sodiumhydroxide, in a suitable solvent such as acetone.

Compound 3 where 4-(X—R⁴) is 4-(SO—R⁴) and 4-(SO₂—R⁴) and R³, R⁴ aredefined as in formula (I), can be obtained by further oxidation of 2where 4-(X—R⁴) is 4-(S—R⁴) with a suitable organic oxidizing agent suchas m-chloroperbenzoic, peracetic acid, etc. in a suitable solvent suchas dichloromethane, or by further oxidation with a suitable inorganicoxidizing agent such as sodium periodate or potassium permangante in asolvent such as water, acetone or acetic acid.

Alternatively, compounds of formula (I) may be prepared according toScheme II from 4. Compound 4, wherein X is O—R⁴ or S—R⁴ and R⁴ is asdefined in formula (I) and R¹ is hydrogen, can be prepared from4-acetoxy or 4-benzoyloxy-azetidin-2-one as described in SyntheticCommunications, 24, 131-135 (1994) whose disclosure is incorporatedherein by reference. Treatment of 4, where R¹ is hydrogen, with asuitable silylating group such as tert-butyldimethylsilyl chloride and asuitable base such as triethylamine in a suitable solvent such astetrahydrofuran gives 4 where R¹ is tert-butyldimethylsilyl. Treatmentof 4, where R¹ is tert-butyldimethylsilyl, with a suitable base such aslithium diisopropylamide in a suitable solvent such as tetrahydrofuranat a suitable temperature such as −50° C., followed by addition to asolution of a suitable azidating reagent such as tosyl azide in asuitable solvent such as tetrahydrofuran, followed by treatment withtrimethylsilyl chloride gives 5 where R¹ is tert-butyldimethylsilyl.Reduction of the azido group in 5, where R¹ is tert-butyldimethylsilyl,with a suitable reducing agent such as hydrogen sulfide in a suitablesolvent such as dichloromethane containing a suitable base such astriethylamine gives 6 where R¹ is tert-butyldimethylsilyl. Treatment of6, where R¹ is tert-butyldimethylsilyl, with a suitable alkylating agentR³Z, where R³ is as defined in formula (I) and Z is a suitable leavinggroup such as chloro, in a suitable solvent such as dimethylformamidecontaining a suitable base such as diisopropylethylamine, gives 2 whereR¹ is tert-butyldimethylsilyl. Treatment of 2, where R¹ istert-butyldimethylsilyl, with a suitable inorganic fluoride, such astetrabutylammonium fluroride, in a suitable solvent such astetrahydrofuran and acetic acid gives 2 where R¹ is hydrogen.

Alternatively, compounds II-5 may be prepared using [2+2] cycloadditionreactions, for example, by following the general procedures described inCama et. al., Tetrahedron Letters, 4233, 1978, whose disclosure isincorporated herein by reference.

Synthetic Chemistry

The invention will now be described by reference to the followingexamples which are merely illustrative and are not to be construed as alimitation of the scope of the present invention. All temperatures aregiven in degrees centigrade, all solvents are highest available purityand all reactions run under anhydrous conditions in an argon atmosphereunless otherwise indicated.

In the Examples, all temperatures are in degrees Centigrade (° C.). Massspectra were performed upon a VG Zab mass spectrometer using fast atombombardment, unless otherwise indicated. ¹H-NMR (hereinafter “NMR”)spectra were recorded at 250 MHz using a Bruker AM 250 or Am 400spectrometer. Multiplicities indicated are: s=singlet, d=doublet,t=triplet, q=quartet, m=multiplet and br indicates a broad signal. Sat.indicates a saturated solution, eq indicates the proportion of a molarequivalent of reagent relative to the principal reactant.

Flash chromatography is run over Merck Silica gel 60 (230-400 mesh).

EXAMPLE 1 Preparation of(3RS,4RS)-4-(Isobutoxy)-3-(triphenylmethylamino)azetidin-2-one

a). 4-(Isobutoxy)azetidin-2-one

A mixture of 4-(benzoyloxy)azetidin-2-one (15.2 g, 80 mmol) in toluene(150 mL) was treated with isobutanol (12 g, 0.16 mol), triethylamine (16g, 0.16 mol) and palladium acetate (3.6 g, 16 mmol), stirred in an icebath for several hours, allowed to warm to RT and stirred for 16 h. Themixture was filtered through Supercel, concentrated and the residue waschromatographed on silica gel eluted with 10-35% ethyl acetate:hexane.Fractions containing the product were combined, concentrated andrechromatographed on silica gel eluted with 10-25% ethyl acetate:hexaneto give the title compound (4.9 g).

b). 1-tert-Butyldimethylsilyl-4-(isobutoxy)azetidin-2-one

A solution of 4-(isobutoxy)azetidin-2-one (4.8 g, 33 mmol) intetrahydrofuran (50 mL) was stirred in an ice bath and treated withtriethylamine (6.6 mL, 66 mmol) followed by dropwise addition of asolution of tert-butyldimethylsilyl chloride (6.5 g, 43 mmol) intetrahydrofuran (15 mL). The mixture was stirred for 5 h in the cold andstored in the refrigerator for 64 h. The mixture was poured into coldwater, extracted with ethyl acetate, and the combined organic phase waswashed with brine, dried (magnesium sulfate), and concentrated. Theresidue was chromatographed on silica gel eluted with 10% ethylacetate:hexane. Fractions containing the product were pooled andconcentrated to give the title compound (6.6 g, 79%).

c).(3RS,4RS)-3-Azido-1-(tert-butyldimethylsilyl)-4-(isobutoxy)azetidin-2-one

A solution of N-(isopropyl)cyclohexylamine (1.5 g, 10 mmol) intetrahydrofuran (18 mL) was cooled to −15° C. and 2 M butyllithium (48mL, 10 mmol) was added dropwise. The reaction mixture was stirred for 40min, the temperature was lowered to −70° C., and the mixture was treateddropwise over 10 min with a solution of1-(tert-butyldimethylsilyl)-4-(isobutoxy)-azetidin-2-one (1.8 g, 7 mmol)in tetrahydrofuran (7 mL). The mixture was stirred for 1 h, transferredto a jacketed addition funnel maintained at −70° C., and added over 40min to a solution of p-toluenesulfonyl azide (1.8 g, 9 mmol) intetrahydrofuran (8 mL) containing hexamethylphosphoramide (2 mL)maintained at −70° C. The reaction was stirred for 1 h and at −50° C.for 4.5 h. The mixture was stirred at −28° C. for 16 h, andtrimethylsilyl chloride (5 mL) was added and the mixture was stirred atRT for 45 min. The mixture was diluted with water and extracted withethyl acetate. The combined organic phases were washed with brine, dried(magnesium sulfate) and concentrated. The residue was chromatographed onsilica gel eluted with 10% ethyl acetate:hexane to give the titlecompound (0.6 g, 30%). MS(ES) m/e 299 [M+H]⁺.

d).(3RS,4RS)-3-Amino-1-(tert-butyldimethylsilyl)-4-(isobutoxy)azetidin-2-one

A solution of(3RS,4RS)-3-azido-1-(tert-butyldimethylsilyl)-4-(isobutoxy)azetidin-2-one(0.2 g, 0.67 mmol) in dichloromethane (15 mL) containing triethylamine(0.07 g, 0.7 mmol) was cooled in an ice bath and hydrogen sulfide wasbubbled through the solution gently for 10 min. The mixture was stirredin the cold for 4 h, concentrated, and then treated with dichloromethaneand concentrated four times to give the title compound.

e).(3RS,4RS)-1-(tert-Butyldimethylsilyl)-4-(isobutoxy)-3-(triphenylmethylamino)azetidin-2-one

A solution of(3RS,4RS)-3-amino-1-(tert-butyldimethylsilyl)-4-(isobutoxy)azetidin-2-one(0.2 g) was dissolved in dimethylformamide (8 mL), cooled in an icebath, and treated with diisopropylethylamine (0.1 mL) followed by tritylchloride (167 mg, 0.6 mmol). The mixture was stirred for 18 h, dilutedwith water (40 mL) and extracted with ethyl acetate. The combinedorganic phase was dried (magnesium sulfate), concentrated, and theresidue was chromatographed on silica gel eluted with 20% ethylacetate:hexane to give the title compound. MS(ES) m/e 515 [M+H]⁺.

f). (3RS,4RS)-4-(Isobutoxy)-3-(triphenylmethylamino)azetidin-2-one

A solution of (3RS,4RS)-1-(tert-butyldimethylsilyl)-4-(isobutoxy)-3-(triphenylmethylamino)azetidin-2-one(160 mg, 0.3 mmol) in tetrahydrofuran (4 mL) was cooled in an ice bathand treated with acetic acid (25 mg, 0.4 mmol) followed by dropwiseaddition of 1 M tetrabutylammonium fluoride (0.6 mL, 0.6 mmol). Themixture was stirred for 20 min and passed through silica gel (10 g)eluted with ethyl acetate. The eluate was concentrated and the residuewas chromatographed on silica gel eluted with 20% ethyl acetate:hexaneto give the title compound. MS(ES) m/e 423 [M+Na]⁺.

EXAMPLE 2 Preparation of (3R,4R)- and(3S,4S)-4-(Isobutoxy)-3-(triphenylmethylamino)azetidin-2-one

(3RS ,4RS)-4-(Isobutoxy)-3-(triphenylmethylamino)azetidin-2-one wasresolved by HPLC (Chiralcel OD, 21×250 mm, 10 mL/min, gradient,A:ethanol B:hexane, 0.5-2.5% A during 20 min, UV detection at 254 nm) toafford the title compounds:

(3R,4R)-4-(isobutoxy)-3-(triphenylmethylamino)azetidin-2-one, t_(R) 35min.

MS(ES) m/e 801 [2 M+H]⁺, and

(3S,4S)-4-(isobutoxy)-3-(triphenylmethylamino)azetidin-2-one, t_(R) 39.9min.

MS(ES) m/e 801 [2 M+H]⁺.

EXAMPLE 3 Preparation of(3R,4R)-4(Isobutylthio)-3-(triphenylmethylamino)azetidin-2-one

A solution of zinc acetate (0.7 g, 3 mmol) in toluene (15 mL) and2-methyl-propanethiol (0.72 g, 8 mmol) was refluxed for 45 min in anapparatus equipped with a Dean-Stark trap to azeotrope water.(3R,4R)-4-(Methylsulfonyl)-3-(triphenylmethylamino)azetidin-2-one (1.5g, 36 mmol) was added and the mixture was heated to 90° C. for 2 h. Themixture was concentrated and the residue was triturated with ethylacetate and the insoluble material was removed by filtration. Thefiltrate was concentrated and the residue was chromatographed on silicagel eluted with 20% ethyl acetate:hexane. The fractions containing theproduct were combined, concentrated to give the title compound. MS(ES)m/e 417 [M+H]⁺.

EXAMPLE 4 Preparation of(3R,4R)-4-(Isobutylsulfonyl)-3-(triphenylmethylamino)azetidin-2-one

A solution of(3R,4R)-4-(isobutylthio)-3-(triphenylmethylamino)azetidin-2-one (50 mg,0.12 mmol) in dichloromethane (2 mL) was cooled in an ice bath andtreated with m-chloroperbenzoic acid (44 mg, 0.25 mmol). The mixture wasstirred for 2.5 h in the cold and partitioned between 5% sodiumcarbonate (5 mL) and dichloromethane. The organic phase was washed with5% sodium carbonate and with brine, dried (magnesium sulfate), filteredand concentrated. The residue was chromatographed on silica gel elutedwith 20% ethyl acetate:hexane and fractions containing the product werepooled and concentrated to give the title compound (25 mg, 47%). MS(ES)m/e 447 [M−H]⁺.

EXAMPLES 5-13

The following compound have been prepared using the procedure of Example3, except substituting isobutanol, propanol, benzyl alcohol, methanol,isobutenol, or octanol for 2-methyl-propanethiol gave:

Example 5: (3S,4S)-4-(Isobutoxy)-3-(triphenylmethylamino)azetidin-2-one:MS(ES) m/e 801 [2 M+H]⁺;

Example 6: (3S,4R)-4-(Isobutoxy)-3-(triphenylmethylamino)azetidin-2-one:MS(ES) m/e 401 [M+H]⁺;

Example 7: (3S,4R)-4-(Propoxy)-3-(triphenylmethylamino)azetidin-2-one:MS(ES) m/e 387 [M+H]⁺;

Example 8: (3S,4S)-4(Propoxy)-3-(triphenylmethylamino)azetidin-2-one:MS(ES) m/e 773 [2M+H]⁺;

Example 9: (3S,4R)-4-(Benzyloxy)-3-(triphenylmethylamino)azetidin-2-one:MS(ES) m/e 435 [M+H]⁺;

Example 10: (3S,4S)-4-(Benzyloxy)-3-(triphenylmethylamino)azetidin-2-one: MS(ES) m/e 435[M+H]⁺;

Example 11: (3S,4R)-4-Methoxy-3-(triphenylmethylamino)azetidin-2-one:MS(ES) m/e 359 [M+H]⁺;

Example 12:(3S,4R)-4-(Isobutenyloxy)-3-(triphenylmethylamino)azetidin-2-one: MS(ES)m/e 399 [M+H]⁺;

Example 13: (3S,4R)-4-Octyloxy-3-(triphenylmethylamino)azetidin-2-one:MS(ES) m/e 457 [M+H]⁺;

EXAMPLES 14-15 Preparation of(3S,4R)-4-Phenoxy-3-(triphenylmethylamino)azetidin-2-and(3S,4S)-4-Phenoxy-3-(triphenylmethylamino)azetidin-2-one

A solution of phenol (0.3 g, 3.2 mmol) in acetone (3 mL) was treatedwith 1 N sodium hydroxide (3.2 mL, 3.2 mmol), stirred 10 min and treateddropwise with a solution of(3R,4R)-4-methylsulfonyl-3-(triphenylmethylamino)azetidin-2-one (1.2 g,3 mmol) in acetone (2 mL). The mixture was stirred for 1.5 h,partitioned between water and diethyl ether, and the combined organicphase was washed with brine, dried (magnesium sulfate), andconcentrated. The residue was chromatographed on silica gel eluted with20% ethyl acetate:hexane to give the title compounds:

(3S,4R)-4-phenoxy-3-(triphenylmethylamino)azetidin-2-one: MS(ES) m/e 443[M+Na]⁺; 421 [M+H]⁺;

(3S,4S)-4-phenoxy-3-(triphenylmethylamino)azetidin-2-one: MS(ES) m/e 841[2 M+H]⁺; 419 [M−H]⁻.

EXAMPLE 16 Preparation of(3S,4R)-3-[[(4-Iodophenyl)diphenylmethyl]amino]4-(isobutoxy)azetidin-2-one

a). (3S,4R)-3-Amino4(isobutoxy)azetidin-2-one para-toluenesulfonate

A solution of para-toluenesulfonic acid hydrate (190 mg) in acetone (20mL) was added to a solution of(3S,4R)-4-isobutoxy-3-(triphenylmethylamino)azetidin-2-one (400 mg, 1mmol) in acetone (20 mL), stirred for 1 h, and concentrated. The residuewas triturated with diethyl ether and the resulting solid isolated byfiltration to give the title compound (180 mg, 55%).

b).(3S,4R)-3-[[(4-Iodophenyl)diphenylmethyl]amino]-4-(isobutoxy)azetidin-2-one

A solution of (3S,4R)-3-amino-4-(isobutoxy)azetidin-2-onepara-toluenesulfonate (180 mg, 0.54 mmol) in acetone (8 mL) containingdiisopropylethylamine ( 1.1 mmol) was treated with a solution of(4-iodophenyl)diphenylmethyl chloride (218 mg, 0.54 mmol), prepared asdescribed by Tschitschibabin, Chem. Ber. 44, 450 (191 1), in acetone (98mL). The solution was stirred for 6 h and partitioned between water anddichloromethane. The combined organic phases were dried (magnesiumsulfate), concentrated, and the residue was chromatographed on silicagel eluted with 15% ethyl acetate:hexane to give the title compound (190mg, 67%). MS(ES) m/e 527 [M+H]⁺.

EXAMPLE 17 Preparation of(3S,4S)-4-[3-(Methoxycarbonyl)propoxy]-3-(triphenylmethylamino)azetidin-2-one

a). (3S,4S)-3-Amino-4-[3-(methoxycarbonyl)propoxy]azetidin-2-one

A solution of methyl4-[(3S,4S)-2-oxo-3-(triphenylmethylamino)azetidin-4-yloxy]but-2-ynoate(75 mg, 0.17 mmol) in absolute ethanol (6 mL) was treated with 10%palladium-on-carbon (35 mg) and stirred under hydrogen overnight. Thecatalyst was removed by filtration and the filtrate concentrated. Theresidual oil was purified on by preparative thin layer chromatography(Whatman, silica gel 60A, 20×20 cm, 1000 um, 20% ethyl acetate:hexane).The origin band contained the title compound (30 mg). MS(ES) m/e 203.0[M+H].

b).(3S,4S)-4-[3-(Methoxycarbonyl)propoxy]-3-(triphenylmethylamino)azetidin-2-one

(3S,4S)-3-Amino-4-[3-(methoxycarbonyl)propoxy]azetidin-2-one (30 mg,0.148 mmol) was dissolved in dry dichloromethane (2 mL) and treated withtriphenylmethyl chloride (41 mg, 0.148 mmol) followed bydiisopropylethylamine (19 mg, 0.148 mmol). The solution was stirred atRT under argon for 5 h, diluted with water and extracted withdichloromethane. The organic phases were combined, washed with water andbrine, dried (magnesium sulfate), filtered, and concentrated. Theresidue was purified by preparative thin layer chromatography (Whatman,silica gel 60A, 20×20 cm, 1000 um, 40% ethyl acetate:hexane) to give thetitle compound (7 mg). MS(ES) m/e 445.2 [M+H].

EXAMPLES 18-21

The following compounds were prepared using the general procedure ofExample 3, except substituting:

Examples 18 and 19: methyl 4-hydroxy-2-butynoate (Zh. Obshch. Khim. 66,106, 1996),

Example 20: 4-hydroxymethyl-2(5 H)-furanone, (J. Chem. Res., Synop. 222,1986),

or

Example 21: 2-(3-pyridylmethyl)-1,3-dithianyl-2-methanol for2-methyl-propanethiol used therein.

Preparation of

EXAMPLE 18: Methyl4-[(3S,4S)-2-oxo-3-(triphenylmethylamino)azetidin-4-yloxy]but-2-ynoate

Methyl4-[(3S,4S)-2-oxo-3-(triphenylmethylamino)azetidin-4-yloxy]but-2-ynoate:

¹H NMR(270 MHz, CDCl₃) δ 3.54 (d, J=17 Hz, 1 H), 3.77 (s, 3 H), 3.83 (d,J=17 Hz, 1 H), 4.08 (s, 1 H), 4.11 (d, J=1.1 Hz, 1 H), 6.39 (s, 1 H),7.19-7.53 (m, 15 H); IR(CHCl₃) 3330, 2240, 1770, 1718, 1477, 1445, 1434,1260, 1194, 1166, 940, 759, 706 cm⁻¹.

EXAMPLE 19: Methyl4-[(3S,4R)-2-oxo-3-(triphenylmethylamino)azetidin-4-yloxy]but-2-ynoate

Methyl4-[(3S,4R)-2-oxo-3-(triphenylmethylamino)azetidin-4-yloxy]but-2-ynoate:

¹H NMR(60 MHz, CDCl₃)δ 2.9 (d, J=9 Hz, 1 H), 3.70 (s, 3 H), 3.60 (d,J=17 Hz, 1H), 3.93 (d, J=17 Hz, 1 H), 4.0-4.3 (m, 2 H), 6.8 (s, 1 H),7.2-7.8 (m, 15 H); IR(CHCl₃) 3350, 1775, 1715 cm⁻¹.

EXAMPLE 20: (3S,4R)-4-[(2(5H)Furanon-4-yl)methoxy]-3-(triphenylmethylamino)azetidin-2-one

(3S,4R)-4-[(2(5H)furanon4-yl)methoxy]-3-(triphenylmethylamino)azetidin-2-one:

¹H NMR(60 MHz, CDCl₃) δ 2.8 (d, J=9 Hz, 1 H), 3.9 (m, 2 H), 4.2 (m, 2H), 4.53 (m, 2 H), 5.87 (m, 1 H), 7.0-8.7 (m, 17 H); IR(CHCl₃) 1785,1755, 1650 cm⁻¹.

EXAMPLE 21: (3S,4R)-4[[2-(3-Pyridylmethyl)-1.3-dithian-2-yl]methoxy]-3-(triphenylmethylamino)azetidin-2-one

(3S,4R)-4-[[2-(3-pyridylmethyl)-1,3-dithian-2-yl]methoxy]-3-(triphenylmethylamino)azetidin-2-one:IR(CHCl₃) 3370, 1775 cm⁻¹.

Using analagous procedures to those indicated above, or as indicated,the following compounds have been syntheized:

EXAMPLE 22:(3S,4S)-4-(Prop-2-ynyloxy)-3-(triphenylmethylamino)azetidin-2-one: J.Chem. Soc. Perkin Trans. I 2268, 1979; EXAMPLE 23:(3R,4R)-4-(Methylsulfonyl)-3-(triphenylmethylamino)azetidin-2-one, J.Chem. Soc. Perkin Trans. I 2268, 1979; EXAMPLE 24:(3S,4R)-4-(Isobutenyloxy)-3-(triphenylmethylamino)azetidin-2-one, J.Chem. Soc. Perkin Trans. I 2268, 1979; EXAMPLE 25:(S)-3-(Triphenylmethylamino)azetidin-2-one: Tetr. Letters 30, 3219,1989. EXAMPLE 26: Using analagous procedures to those noted therein, theracemic mixture of (RS)-3-(Triphenylmethylamino)azetidin-2-one may beproduced.

Methods of Treatment

The compounds of Formula (I) or pharmaceutically acceptable saltsthereof can be used in the manufacture of a medicament for theprophylactic or therapeutic treatment of an inflammatory disease statein a mammal, preferably a human.

Inhibition of COA-IT and the simultaneous reduction of PAF, freearachidonic acid and eicosanoid release from inflammatory cellsaccording to this invention is of therapeutic benefit in a broad rangeof diseases or disorders. The invention herein is therefore useful totreat such disease states both in humans and in other mammals.

Inhibition of CoA-IT by the compounds of Formula (I) is an effectivemeans for simultaneously reducing PAF, free arachidonic acid andeicosanoids produced in inflammatory cells. The therapeutic utility ofblocking lipid mediator generation has been recognized for many years.For example, inhibitors of cyclooxygenase, such as aspirin,indomethacin, acetaminophen and ibuprofen, have demonstrated broadtherapeutic utilities. CoA-IT inhibitors inhibit cyclooxygenaseproducts. Another class of inhibitors which are used in a broad range ofinflammatory disorders are the corticosteroids. Corticosteroids act in avariety of ways, e.g. to induce inflammatory cells to produce proteinswhich inhibit free arachidonic acid release or to down regulate PLA₂mRNA formation. Both 14 kDa PLA2 inhibitors and CoA-IT inhibitors blockthe release of free arachidonic acid. Inhibitors of 5-lipoxygenase blockthe production of leukotrienes and leukotriene antagonists prevent thebioactions of leukotrienes. Recent studies indicate that both will havebroad therapeutic utilities. Both 14 kDa PLA2 inhibitors and CoA-ITinhibitors block the production of leukotrienes. Inhibitors ofphospholipase A₂ block the release of free arachidonic acid and theformation of lyso PAF (the immediate precursor of PAF). PLA₂ inhibitorsare recognized to have broad therapeutic utilities. It does not ,however, follow that the disease states noted above must in fact becaused by altered CoA-IT activity. Thus, the disease state itself maynot be directly mediated by CoA-IT activity. It only follows that CoA-ITactivity is required for the continued expression of symptoms of thedisease state and that CoA-IT inhibitors will be beneficial against thesymptoms of these disease states.

Recognition that CoA-IT inhibitors reduce PAF production has a number oftherapeutic implications. PAF itself has been implicated as beinginvolved in a number of medical conditions. Thus in circulatory shock,which is characterised by systemic hypotension, pulmonary hypertensionand increased lung vascular permeability, the symptoms can be mimickedby infusion of PAF. This coupled with evidence showing that circulatingPAF levels are increased by endotoxin infusion indicate that PAF is aprime mediator in certain forms of shock.

Intravenous infusion of PAF at doses of 20-200 pmol kg<−1>min<−1> intorats has been reported to result in the formation of extensivehaemorrhagic erosions in the gastric mucosa. Thus PAF is the most potentgastric ulcerogen yet described whose endogenous release may underlie orcontribute to certain forms of gastric ulceration. Psoriasis is aninflammatory and proliferative disease characterised by skin lesions.PAF is pro-inflammatory and has been isolated from lesioned scale ofpsoriatic patients indicating PAF has a role is the disease ofpsoriasis. And finally, increasing evidence supports a potentialpatho-physiological role for PAF in cardiovascular disease. Thus recentstudies in angina patients show PAF is released during atrial pacing.Intracoronary injection of PAF in pigs induces a prolonged decrease incoronary flow and, in guines pig hearts, it induces regional shuntingand ischaemia. In addition PAF has been shown to initiate thrombusformation in a mesenteric artery preparation, both when administeredexogenously and when released endogenously. More recently PAF has beenshown to play a role in brain ischaemia induced in animal models ofstroke. Thus the compounds of the invention, by virtue of their abilityto antagonise CoA-IT thus block the production of PAF, free arachidonicacid and its metabolites, are likely to be of value in the treatment ofany of the above conditions.

Treatment of disease states caused by these lipid inflammatory mediatorsi.e., arachidonate, eicosanoids and PAF, include certain cardiovasculardisorders such as but not limited to, myocardial infarction, stroke,circulatory shock, or hypotension, ischemia, reperfusion injury;inflammatory diseases such as, but not limited to, arthritis,inflammatory bowel disease, Crohn's disease, or ulcerative colitis;respiratory diseases such as but not limited to, asthma, or adultrespiratory distress syndrome; analphylaxis, shock, such as but notlimited to endotoxic shock; topical disesases, such as but not limitedto actinic keratosis, psoriasis, or contact dermatitis; or pyresis.

In order to use a compound of formula (I) or a pharmaceuticallyacceptable salt thereof in therapy, it will normally be formulated intoa pharmaceutical composition in accordance with standard pharmaceuticalpractice. This invention, therefore, also relates to a pharmaceuticalcomposition comprising an effective, non-toxic amount of a compound offormula (I) and a pharmaceutically acceptable carrier or diluent.

Compounds of formula (I), pharmaceutically acceptable salts thereof andpharmaceutical compositions incorporating such may conveniently beadministered by any of the routes conventionally used for drugadministration, for instance, orally, topically, parenterally or byinhalation. The compounds of formula (I) may be administered inconventional dosage forms prepared by combining a compound of formula(I) with standard pharmaceutical carriers according to conventionalprocedures. Such pharmaceutically acceptable carriers or diluents andmethods of making are well known to those of skill in the art, andreference can be found in such texts as Remington's PharmaceuticalSciences, 18th Ed., Alfonso R. Genarao, Ed., 1990, Mack Publishing Co.and the Handbook of Pharmaceutical Excipents, APhA Publications, 1986.

The compounds of formula (I) may also be administered in conventionaldosages in combination with known second therapeutically activecompounds, such as steroids or NSAID's for instance. These proceduresmay involve mixing, granulating and compressing or dissolving theingredients as appropriate to the desired preparation. It will beappreciated that the form and character of the pharmaceuticallyacceptable carrier or diluent is dictated by the amount of activeingredient with which it is to be combined, the route of administrationand other well-known variables. The carrier(s) must be “acceptable” inthe sense of being compatible with the other ingredients of theformulation and not deleterious to the recipient thereof.

The pharmaceutical carrier employed may be, for example, either a solidor liquid. Exemplary of solid carriers are lactose, terra alba, sucrose,talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acidand the like. Exemplary of liquid carriers are syrup, peanut oil, oliveoil, water and the like. Similarly, the carrier or diluent may includetime delay material well known to the art, such as glycerylmono-stearate or glyceryl distearate alone or with a wax.

A wide variety of pharmaceutical forms can be employed. Thus, if a solidcarrier is used, the preparation can be tableted, placed in a hardgelatin capsule in powder or pellet form or in the form of a troche orlozenge. The amount of solid carrier will vary widely but preferablywill be from about 25 mg. to about 1 g. When a liquid carrier is used,the preparation will be in the form of a syrup, emulsion, soft gelatincapsule, sterile injectable liquid such as an ampule or nonaqueousliquid suspension.

Compounds of formula (I) may be administered topically, that is bynon-systemic administration. This includes the application of a compoundof formula (I) externally to the epidermis or the buccal cavity and theinstillation of such a compound into the ear, eye and nose, such thatthe compound does not significantly enter the blood stream. In contrast,systemic administration refers to oral, intravenous, intraperitoneal andintramuscular administration.

Formulations suitable for topical administration include liquid orsemi-liquid preparations suitable for penetration through the skin tothe site of inflammation such as liniments, lotions, creams, ointmentsor pastes, and drops suitable for administration to the eye, ear ornose. The active ingredient may comprise, for topical administration,from 0.001% to 10% w/w, for instance from 1% to 2% by weight of theformulation. It may however comprise as much as 10% w/w but preferablywill comprise less than 5% w/w, more preferably from 0.1% to 1% w/w ofthe formulation.

Lotions according to the present invention include those suitable forapplication to the skin or eye. An eye lotion may comprise a sterileaqueous solution optionally containing a bactericide and may be preparedby methods similar to those for the preparation of drops. Lotions orliniments for application to the skin may also include an agent tohasten drying and to cool the skin, such as an alcohol or acetone,and/or a moisturizer such as glycerol or an oil such as castor oil orarachis oil.

Creams, ointments or pastes according to the present invention aresemi-solid formulations of the active ingredient for externalapplication. They may be made by mixing the active ingredient infinely-divided or powdered form, alone or in solution or suspension inan aqueous or non-aqueous fluid, with the aid of suitable machinery,with a greasy or non-greasy base. The base may comprise hydrocarbonssuch as hard, soft or liquid paraffin, glycerol, beeswax, a metallicsoap; a mucilage; an oil of natural origin such as almond, corn,arachis, castor or olive oil; wool fat or its derivatives or a fattyacid such as steric or oleic acid together with an alcohol such aspropylene glycol or a macrogel. The formulation may incorporate anysuitable surface active agent such as an anionic, cationic or non-ionicsurfactant such as a sorbitan esteror a polyoxyethylene derivativethereof. Suspending agents such as natural gums, cellulose derivativesor inorganic materials such as silicaceous silicas, and otheringredients such as lanolin, may also be included.

Drops according to the present invention may comprise sterile aqueous oroily solutions or suspensions and may be prepared by dissolving theactive ingredient in a suitable aqueous solution of a bactericidaland/or fungicidal agent and/or any other suitable preservative, andpreferably including a surface active agent. The resulting solution maythen be clarified by filtration, transferred to a suitable containerwhich is then sealed and sterilized by autoclaving or maintaining at98-100 ° C. for half an hour. Alternatively, the solution may besterilized by filtration and transferred to the container by an aseptictechnique. Examples of bactericidal and fungicidal agents suitable forinclusion in the drops are phenylmercuric nitrate or acetate (0.002%),benzalkonium chloride (0.01%) and chlorhexidine acetate (0.01%).Suitable solvents for the preparation of an oily solution includeglycerol, diluted alcohol and propylene glycol.

Each dosage unit for oral administration contains preferably from 1 to250 mg (and for parenteral administration contains preferably from 0.1to 25 mg) of a compound of the structure (I) or a pharmaceuticallyacceptable salt thereof calculated as the free base.

The pharmaceutically acceptable compounds of the invention will normallybe administered to a subject in a daily dosage regimen. For an adultpatient this may be, for example, an oral dose of between 1 mg and 500mg, preferably between 1 mg and 250 mg, or an intravenous, subcutaneous,or intramuscular dose of between 0.1 mg and 100 mg, preferably between0.1 mg and 25 mg, of the compound of the Formula (I) or apharmaceutically acceptable salt thereof calculated as the free base,the compound being administered from 1 to 4 times per day.

The choice of form for administration, as well as effective dosages,will vary depending, inter alia, on the condition being treated. Thechoice of mode of administration and dosage is within the skill of theart.

Biological Methods:

To determine activity of the compounds of Formula (I) various cellularassays can be used to determine in vitro activity. Additionally, variousclassical in vivo acute inflammatory models which have some aspect oftheir etilogy to elevated eicosanoid levels can be employed, such as thepaw edema model, mouse zymosan peritonitis, reverse Arthus pleurisy orvarious skin inflammation assays which are described in Lewis et al.,Experimental Models of Inflammation, in the Handbook of Inflammation,Vol. 5, Bonta Ed., Elsevier Science Publishers, N.Y. (1985) whosedisclosure is herein incorporated by reference. The TPA induced earedema model (mouse) is described herein as well. These classical modelsof inflammation will reflect the drug's ability to alter an inflammatoryresponse but cannot address the specificity of drug action. These modelshave been traditionally designed as non steriod antiinflammatory drugsensitive pharmacological screens and it is important to utilize modelswhich can differentiate PLA2 and CoA-IT inhibitors from NSAIDS.

Cell-free and Cellular Assessment of Inhibitors

Described herein is an in vitro assay for CoA-IT enzyme activity. Acellular assay for PAF production is also described herein.

Inflammatory Responses in vivo

The ability of compounds that inhibit CoA-IT to affect in vivoinflammatory responses may be assessed. Inflammatory responses areinduced in the mouse ear by the topical application of apro-inflammatory agent, such as 12-O-tetradecanoyl-phorbol 13-acetate.This produces an edematous response, as measured by increases in earthickness, as well as increased inflammatory cellular infiltrate, asmeasured by increases in myeloperoxidase activity (as described in themethods). To further validate the mechanism of action inflammationinduced by the direct adminstration of arachidonic acid can be used. Inthis case compounds altering arachidonic acid mobilization or liberationshould be with our effect.

In Vitro Assays

Assay: CoA-IT Activity

The following is a method to measure CoA-IT activity and the effects ofcompounds on CoA-IT activity. The assay is based upon mixing cellularmaterial containing CoA-IT activity with a stable lyso phospholipid suchas 1-alkyl-2-acyl-GPC and measuring the production of phospholipidproduct such as 1-alkyl-2-acyl-GPC occurring in the absence of added CoAor CoA-fatty acids.

Cell Preparation

Any inflammatory cell that contains high levels of COA-IT activity canbe used, such as neutrophils, macrophages or cell lines such as U937cells. U937 cells were obtained from American Type Culture Collectionand grown in RPMI- 1640 media (Gibco, Grand Island, N.Y.) supplementedwith 10% fetal bovine serum (Hyclone, Logan, Utah) at 37° C., 5% CO₂.Cells were grown without differentiation (basal state) by any agent,such as dimethyl sulfoxide. As used herein, “inflammatory cells”include, but are not limited to neutrophils, macrophages, monocytes,lymphocytes, eosinophils, basophils, and mast cells.

Microsomal preparation

Microsomes were prepared using standard techniques. In this case, cellswere washed with a buffer of 250 mM sucrose, 10 mM Tris, 1 mM EGTA, 1 mMMgCl₂, pH 7.4 and ruptured by N₂ cavitation (750 psi, 10 minutes). Theruptured cells were centrifuged 1000×g, 5 minutes. The resultingsupernatant was centrifuged at 20,000×g, ˜20 minutes. Microsomes wereprepared from this supernatant by centrifugation at 100,000×g, 60minutes. The resulting pellet was washed once with assay buffer (150 mMNaCl, 10 mM Na₂KPO₄, 1 mM EGTA, pH 7.4), recentrifuged and the pelletresuspended in assay buffer (4-20 mg protein/ml) and was stored at −80°C. until assayed.

CoA-IT activity

CoA-lT activity was measured in 1.5 ml centrifuge tubes in a totalvolume of 100 ul. Microsomes were diluted in assay buffer to the desiredprotein concentration (6-20 ug/tube). The reaction was initiated byaddition of [³H]1-alkyl-2-lyso-sn-glycero-3-phosphocholine (GPC) (˜0.1uCi/tube) and 1 μM final cold 1-alkyl-2-lyso-GPC in assay buffer with0.25 mg/ml fatty acid-poor bovine serum albumin (BSA) (Calbiochem, LaJolla, Calif.). [³H]1-alkyl-2-lyso-GPC, approximately 50 Ci/mmol, wasfrom NEN-Dupont (Boston, Mass.) and cold 1-alkyl-2-lyso-GPC was fromBiomol (Plymouth Meeting, Pa.). Microsomes were pretreated with desiredagents for the desired time (10 minutes) before the addition of[³H]1-alkyl-2-lyso-GPC. The reaction was run for the desired time (10minutes) at 37° C. The reaction was stopped and the lipids extracted byaddition of 100 ul of chloroform:methanol (1:2, v/v) followed by 100 ulof chloroform and 100 ul of 1 M KCl. The samples were vortexed andcentrifuged at high speed in a microfuge for 2-3 minutes. An aliquot ofthe chloroform-extracted materials were separated, usually by TLC inchloroform/methanol/acetic acid/water (50:25:8:4, v/v), visualized byradioscanning (Bioscan) and the product, [³H]1-alkyl-2-acyl-GPC, wasscraped and quantified by liquid scintillation spectroscopy. With thisTLC system, the synthetic standards of 1-alkyl-2-lyso-GPC and1-alkyl-2-acyl-GPC were well separated, with Rf values of approximately0.25 and 0.65, respectively. Other methods can be used to separatesubstrate from product, including but not limited to columnchromatography, affinity chromatography and post reactionderivitization.

Protein concentration were assessed using the protein assay reagentsfrom Bio-Rad (Richmond, Calif.).

Results

A variety of compounds have been tested in this assay to determine itsselectivity and inability to detect trivial, non-selective inhibitors.Inhibitors of 5-lipoxygenase (5-LO) and cyclooxygenase (CO), such asindomethicin, naproxen,6-(4′-fluorophenyl)-5-(4-pyridyl)-2,3-dihydroimidzo-[2,1-b]thiazole and6-(4′-fluorophenyl)-5-(4-pyridyl)2,3-dihydroimidzo-[2,1-b]thiazole-dioxidehad no effect on CoA-IT activity at concentrations up to 100 μM. Theanti-oxidant BHT also has no effect at concentrations up to 100 μM.Compounds which complex with phospholipids and inhibit PLA₂ activity,such as quinacrine and aristolochic acid have no effect on CoA-ITactivity at concentrations up to 500 μM. Doxepine, a compound reportedto inhibit PAF release did not inhibit CoA-IT at concentrations up to100 μM. Sodium diclofenac, reported to decrease leukotriene productionby altering arachidonic acid metabolism, had no effect on CoA-lTactivity at concentrations up to 500 μM. These results show that theassay for CoA-IT activity is sensitive and selective.

FIG. 1 demonstrates the inhibition of CoA-IT activity in a timedependent manner using a representative compound of Formula (I), Example24, (3S,4R)-4-(isobutenyloxy)-3-(triphenylmethylamino)azetidin-2-one.

Other representative compounds of Formula (I) which inhibited CoA-ITactivity, as a time-dependent inhibitor, in the microsomal CoA-IT assaydescribed above [generally at an IC₅₀ of <50 μM or less, at a 10 minpreincubation time] are:

(3RS,4RS)-4-(Isobutoxy)-3-(triphenylmethylamino)azetidin-2-one

(3R,4R)-4-(Isobutoxy)-3-(triphenylmethylamino)azetidin-2-one

(3S,4S)-4-(Isobutoxy)-3-(triphenylmethylamino)azetidin-2-one

(3R,4R)-4-(Isobutylthio)-3-(triphenylmethylamino)azetidin-2-one

(3R,4R)-4-(Isobutylsulfony)-3-(triphenylmethylamino)azetidin-2-one

(3S,4R)-4-(Isobutoxy)-3-(triphenylmethylamino)azetidin-2-one

(3S,4R)-4-(Propoxy)-3-(triphenylmethylamino)azetidin-2-one

(3S,4S)-4-(Propoxy)-3-(triphenylmethylamino)azetidin-2-one

(3S,4R)-4-(Benzyloxy)-3-(triphenylmethylamino)azetidin-2-one

(3S,4S)-4-(Benzyloxy)-3-(triphenylmethylamino)azetidin-2-one

(3S,4R)-4-Methoxy-3-(triphenylmethylamino)azetidin-2-one

(3S,4R)-4-(Isobutenyloxy)-3-(triphenylmethylamino)azetidin-2-one

(3S,4R)-4-Octyloxy-3-(triphenylmethylamino)azetidin-2-one

(3S,4R)-4-Phenoxy-3-(triphenylmethylamino)azetidin-2-one

(3S,4S)-4-Phenoxy-3-(triphenylmethylamino)azetidin-2-one

(3S,4R)-3-[[(4-Iodophenyl)diphenylmethyl]amino]-4-(isobutoxy)azetidin-2-one

(3S,4R)-4-[[2-(Pyrid-3-yl)-1,3-dithian-2-yl]methoxy]-3-(triphenylmethylamino)azetidin-2-one

(3S,4S)-4-(Prop-2-ynyloxy)-3-(triphenylmethylamino)azetidin-2-one

Methyl4-[(3S,4S)-2-oxo-3-(triphenylmethylamino)azetidin4-yloxy]but-2-ynoate

Methyl4-[(3S,4R)-2-oxo-3-(triphenylmethylamino)azetidin-4-yloxy]but-2-ynoate

(3S,4R)-4-[(2(5H)Furanon-4-yl)methoxy]-3-(triphenylmethylamino)azetidin-2-one

(S)-3-(Triphenylmethylamino)azetidin-2-one

(RS)-3-(Triphenylmethylamino)azetidin-2-one

(3S,4S)-4-[3-(Methoxycarbonyl)propoxy]-3-(triphenylmethylamino)azetidin-2-one

(3R,4R)-4-(Methylsulfonyl)-3-(triphenylmethylamino)azetidin-2-one.

The following compound was found active at either increased uM or longerpretreatment time:

(3S,4S)-4-[3-(Methoxycarbonyl)propoxy]-3-(triphenylmethylamino)azetidin-2-one

Assay Platelet-activating Factor (PAF)

Preparation of Human Neutrophils

Human neutrophils are obtained in the laboratory using three differentmethods. One method uses leukophoresis packs from normal humans andneutrophils are isolated using the histopaque- 1077 technique. The bloodis centrifuged at 300×g for 10 minutes. The cell pellets are resuspendedin PBS composed of 137 mM NaCl, 8.8 mM Na₂HPO₄, 1.5 mM KH₂PO₄, 2.7 mMKCl (Dulbecco's Gibco Laboratories, Long Island, N.Y.) and layered overhistopaque-1077 (Sigma, St. Louis, Mo.). The pellets are collected aftercentrifugation (300×g for 30 minutes) and washed once in PBS. The cellpellets are exposed briefly to deionized water to lyse any erythrocytes.The remaining cells are collected by centrifugation, suspended in PBS,counted and identified after cytospinning and staining. The finalleukocyte preparation will be of greater than 95% purity and viability.

The second method isolates human neutrophils from fresh heparinizednormal blood using the Histopaque-1077 technique. The blood is layeredover Histopaque-1077 (Sigma, St. Louis Mo.) and centrifuged at 400×g for30 minutes. The cell pellets are resuspended in 35 ml of PBS and 12 mlof 6% Dextran, followed by Dextran sedimentation at room temperature for45 minutes. The upper layer is collected and further centrifugated for10 minutes at 1000 rpm. The cell pellets are exposed briefly todeionized water to lyse erythrocytes. The remaining cells are collectedby centrifugation, suspended in PBS, counted and identified aftercytospinning and staining. The final leukocyte preparation will be ofgreater than 95% purity and viability.

The third method isolates human neutrophils from freshly drawnheparinized normal blood using the Percoll technique. The blood is firsttreated with 6% Dextran at room temperature for a 1 hour sedmination.The upper layers of plasma are collected and centrifuged at 400×g for 10minutes. The cell pellets are resuspended in Percoll 1.070 g/mlsupplemented with 5% fetal bovine serumand layered on discontinuousgradients (1.080, 1.085, 1.090,1.095 g/ml) followed by centrifugation at400×g for 45 minutes. The neutrophils are collected from interfaces of1;080 and 1.085 and the 1.085 and 1.090 Percoll densities, followed by acentrifugation at 400×g for 45 minutes. The neutrophils are suspended inPBS, counted and identified after cytospinning and staining. The finalleukocyte preparation will be of greater than 95% purity and viability.

There should be no difference noted in the response of the neutrophilsnor in the effects of test compounds in neutrophils isolated by thethree different techniques.

Treatment of Human Neutrophils

Neutrophils were suspended in PBS at concentrations of 5 to 20×10⁶ cellsper ml. Cells were added to test tubes and treated with the desiredcompounds for 5 to 10 minutes, then challenged with calcium ionophoreA23187, 2 μM and 20-30 μCi of [³H] acetic acid (NEN-Dupont, Boston,Mass.), or the vehicle of PBS with 0.25-1 mg/ml. After 5 to 20 minutes,the reactions were terminated by addition of an equal volume ofchloroform:methanol (1:2, v/v) to the samples and the lipids wereextracted by addition of equal volumes of chloroform and distilledwater. The samples were vortexed and centrifuged at high speed and thechloroform layer removed to a clean tube.

Assay for PAF

The chloroform from each tube was evaporated to dryness and the materialsuspended in a small volume of chloroform or chloroform:methanol (25-100μl) and the total material spotted on a Silica TLC plate. The plateswere developed in chloroform/methanol/acetic acid/water (50:25:8:4, v/v)visualized by radioscanning (Bioscan) and the product, [³H]PAF, wasscraped and quantified by liquid scintillation spectroscopy. With thisTLC system, the Rf value for a synthetic standard of PAF isapproximately 0.33.

FIG. 2, demonstrates that a representative compound of Formula 1,Example 24(3S,4R)-4-(Isobutenyloxy)-3-(triphenylmethylamino)azetidin-2-one blocksPAF production in neutrophils.

Assay: Measurement of Stimulated Eicosanoid Release by Human Monocytes.

Human Monocyte Isolation. Leukocyte-rich leukopaks obtained fromBiological Specialties (Lansdale, Pa.) were collected from malevolunteers who were not taking anti-inflammatory drugs. Leukopaks werecentrifuged (90×g for 15 min) twice to remove the platelet-rich plasma.The cell pellet was washed by centrifugation and resuspended in HBSSwithout Ca²⁺ or Mg²⁺. Histopaque 1077 was layered under the cellsuspension and centrifuged at 400×g for 30 min to obtain the buffy coat.The interfacial buffy coat, containing monocytes and lymphocytes, wasremoved and saved. The buffy coat was washed twice with HBSS withoutCa²⁺ or Mg²⁺ by centrifugation. The cell pellet (4-6×10⁸ cells/30mls)was resuspended in iso-osmotic media (RPMI-1640, 10% heat inactivatedfetal bovine serum (FBS), 0.2 mM L-glutamine, 2.5 mM HEPES) and layeredover an equal volume of 46% Percol mixture (10X PBS/Percol; 9.25/0.75)and 54% iso-osmotic media and centrifuged for 30 min at 1000×g (Marshalland Roshak. Biochem. Cell Biol. 71: 331-339, 1993). The monocytepopulation located at the interface of the Percoll gradient was removedand washed twice in HBSS without Ca²⁺ or Mg²⁺. This resulted in agreater than 85-90% pure monocyte population as assessed by differentialstaining.

Measurement of Stimuli-Induced Eicosanoid Release. Monocytes (5×10⁶/ml)were incubated as a suspension in serum-free RPMI-1640 medium containingthe vehicle DMSO (<1%) or drug for 30 min at 27° C. after which vehicleor stimuli was added for the indicated time. The stimulating agent issolubilized in DMSO and appropriate vehicle controls were included inall experiments. The amount of stimuli was chosen from the linearportion of a concentration versus product curve usually representing60-80% maximal stimulation over the indicated incubation time at 37° C.(A23187, 1 μM. 15 min). The reaction was terminated by reduction of pHthrough addition of citric acid and centrifugation (10 min, 400×g. 4°C.). Cell viability was monitored before and after experiments usingtrypan blue exclusion. The cell-free media was decanted and stored al.−70° C. until analyzed. Prostaglandin E₂ and LTC₄ were directly measuredin cell-free media using enzyme immunoassay (EIA) kits purchased fromCaymen Chemical Co. (Ann Arbor, Mich.). Sample or standard dilutionswere made with appropriate media and analyzed in triplicate. Resultswere obtained by extrapolation from a standard curve prepared in themedia and expressed as pg or ng/ml of sample.

FIGS. 3 and 4 demonstrate that a representative compound of Formula (I),(3S,4R)-4-(isobutenyloxy)-3-(triphenylmethylamino)azetidin-2-one.Example 24 demonstrated positive activity in this assay. These Figuresshows that this compound through its CoA-IT inhibition blocked theproduction of leukotriene and prostaglandin, and that such inhibitionwas dependent upon the pre-treatment time.

In Vivo Assays

Assay: Assay (Method) for TPA-induced Inflammation

Animals:

Male Balb/c inbred mice were obtained from Charle River BreedingLaboratories (Kingston, N.Y.). Within a single experiment mice (22-25 g)were age-matched. These in vivo experiments typically involved use of5-6 animals/group.

TPA-induced Mouse Ear Inflammation:

Assay of Ear-Edema:

TPA (12-O-tetradecanoylphorbol 13-acetate) (Sigma Chemical Company) inacetone (4 mg/20 ml) was applied to the inner and outer surfaces of theleft ear of BALB/c male mice. The thickness of both ears was thenmeasured with a dial micrometer (Mitutoyo, Japan) at both 2 and 4 hoursafter treatment, and the data expressed as the change in thickness (10⁻³cm) between treated and untreated ears. The application of acetone didnot cause an edematous response; therefore, the difference in earthickness represented the response to the TPA. After measuring theedema, the inflammed left ears were removed and stored at −70° C. untilthey were assayed for MPO (myeloperoxidase) activity where appropriate.

Assay of Myeloperoxidase (MPO) in Inflamed Ear Tissue:

On the day of the assay, partially thawed ear tissues were minced andthen homogenized (10% w/v) with a Tissumnizer homogenizer (Tekmar Co.)in 50 mM phosphate buffer (pH 6) containing 0.5% HTAB. The tissuehomogenates were taken through three cycles of freeze-thaw, followed bybrief sonication (10 sec). The method of Bradley et al. was used withmodifications as described. The appearance of a colored product from theMPO-dependent reaction of o-dianisidine (0.167 mg/ml; Sigma) andhydrogen peroxide (0.0005%; Sigma) was measured spectrophotometricallyat 460 nm. Supernatant MPO activity was quantified kinetically (changein absorbance measured over 3 min, sampled at 15-sec intervals) using aBeckman DU-7 spectrophotometer and a Kinetics Analysis package (BeckmanInstruments, Inc.). One unit of MPO activity is defined as thatdegrading one micromole of peroxide per minute at 25° C.

Statistics:

Statistical analysis was done using Student's “t” test. The ED₅₀ arevalues which cause a 50% inhibition of the inflammatory response and arecalculated by regression analysis of the dose response data.

As shown below in Table 1, a representative compound of Formula (I),(3S,4R)-4-(isobutenyloxy)-3-(triphenylmethylamino)azetidin-2-one,Example 24 demonstrated positive activity in this assay.

TABLE 1 Effects of topically administered compounds on phorbol ester-induced inflammation in mouse ear. Edema @ 4 hr MPO response CompoundED₅₀ (mg/ear) ED₅₀ (mg/ear) CoA-IT 0.49 0.48 Example 24 Steroid 0.060.03 dexamethasone

Balb/c mice were given PMA followed by the test compound. The edematousresponse was measured using a thickness gauge 4 hrs post PMAapplication. The animals were sacrificed and the inflamed ears harvestedand MPO extracted and assayed spectrophotometrically as described inmethods. Both compounds produced effects significantly different fromvehicle treatment.

The positive activity of the compounds of Formula (I) in this animalmodel demonstrate a clear utility in the treatment of topicallyadministered diseases associated with inflammation as noted herein suchas, but not limited to, inflammatory bowel disease, contact dermatoses,actinic keratosis, psoriasis, or conjunctivitis.

As used herein, various abbreviations and explanations are as follows:[³H], a molecule that contains tritium atoms, a radioactive isotope;A23187, a compound that allows free entry of calcium into a cell; AA,arachidonic acid, 5-8-11-14 eicosatetraenoic acid; arachidonate,arachidonic acid contained within a phospholipid; free arachidonic acid,arachidonic acid that is not contained within a phospholipid;[²H₈]arachidonic acid, the form of arachidonic acid labeled with 8deuterium atoms, a stable isotope; 1-alkyl, 1-O-alkyl; 1-alkenyl,1-O-alk-1′-enyl; BSA, bovine serum albumin; CoA, coenzyme A; CoA-IT,CoA-independent transacylase; COX, cyclooxygenase; DTT, dithiothreitol;EGTA, [ethylenebis(oxyethylenenitrilo)]tetra acetic acid, a calciumchelator; GPC, sn-glycero-3-phosphocholine; EDTA, a metal ion chelator;GPE, sn-glycero-3-phosphoethanolamine; GC/MS, gas chromatography andmass spectrometry; 5HETE, 5(S)-hydroxyeicosa-6,8,11,14-tetraenoic acid;15HETE, 15(S)-hydroxyeicosa-5,8, 11,13-tetraenoic acid; HL-60, AmericanType Tissue Culture designated cell line similar to a monocyte; 5LO,5-lipoxygenase; LTB₄, leukotriene B₄; LTC₄, leukotriene C₄; LTD₄,leukotriene D₄; lyso PAF, 1-alkyl-2-lyso-GPC, lyso platelet-activatingfactor; PLA₂, phospholipase A₂; PBS, phosphate buffered saline; PAF,platelet activating factor, 1-alkyl-2-acetyl-GPC; PL, phospholipid; PC,phosphatidylcholine; PE, phosphatidylethanolamine, PI,phosphatidylinositol; PMN, polymorphonuclear neutrophilic cell,neutrophil; PS phosphatidylserine; Rf, the distance a compound travelsas a fraction of the solvent front; TLC, thin layer chromatography;U937, American Type Tissue Culture designated cell line similar to amonocyte.

All publications, including but not limited to patents and patentapplications, cited in this specification are herein incorporated byreference as if each individual publication were specifically andindividually indicated to be incorporated by reference herein as thoughfully set forth.

The above description fully discloses the invention including preferredembodiments thereof. Modifications and improvements of the embodimentsspecifically disclosed herein are within the scope of the followingclaims. Without further elaboration, it is believed that one skilled inthe art can, using the preceding description, utilize the presentinvention to its fullest extent. Therefore, the Examples herein are tobe construed as merely illustrative and not a limitation of the scope ofthe present invention in any way. The embodiments of the invention inwhich an exclusive property or privilege is claimed are defined asfollows.

What is claimed is:
 1. A method for treating disease or disordersmediated by a lipid inflammatory mediator selected from arachidonricacid, its metabolites or platelet activating factor (PAF), which methodcomprising administering to a mammal in need thereof an effectiveCoenzyme A independent transacylase (CoA-IT) inhibiting amount of acompound of the Formula:

wherein Y is NH; X is O or S(O)m; m is 0 or an integer having a value of1, or 2; R₃ is optionally substituted triphenylmethyl; R₄ is optionallysubstituted C₁₋₁₀ alkyl, (CR₁₀R₂₀)_(n)C(R₁₀)=C(R₇)₂, or(CR₁₀R₂₀)n—C≡C—R₅; n is an integer having a value of 1 to 4; R₁₀ and R₂₀are independently hydrogen or C₁₋₄ alkyl; R₅ is hydrogen, alkyl, aryl,arylalkyl, heteroaryl, heteroarylalkyl, C(O)₂R₆, or C(O)R₆ wherein thealkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl moieties may beoptionally substituted; R₆ is C₁₋₁₀ alkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, heterocyclic, or heterocyclicalkyl, wherein the alkyl,aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclic, orheterocyclicalkyl moieties may be optionally substituted; R₇ isindependently hydrogen, C₁₋₁₀ alkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, heterocyclic, or heterocyclicalkyl, wherein the alkyl,aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclic, orheterocyclicalkyl moieties may be optionally substituted; or apharmaceutically acceptable salt thereof.
 2. The method according toclaim 1 wherein the disease or disorder is allergic rhinitis, asthma,myocardial infarction, stroke, circulatory shock, hypotension, ischemia,reperfusion injury, arthritis, inflammatory bowel disease, Crohn'sdisease, ulcerative colitis, asthma, adult respiratory distresssyndrome, analphylaxis, shock, endotoxic shock, actinic keratosis,psoriasis, contact dermatitis, pyresis, or any other disease, disorderor syndrome mediated in at least part by lipid inflammatory mediators.3. The method according to claim 1 wherein R₄ is an optionallysubstituted C₁₋₁₀ alkyl, or alkenyl.
 4. The method according to claim 3wherein the alkyl is substituted by a ketal or thioketal.
 5. The methodaccording to claim 1 wherein R₅ is hydrogen, C(O)₂R₆ or a heteroarylring.
 6. The method according to claim 5 wherein the heteroaryl ring isa pyridyl ring.
 7. The method according to claim 1 wherein the compound,or a pharmaceutically accetable salt thereof, is:(3RS,4RS)-4-(Isobutoxy)-3-(triphenylmethylamino)azetidin-2-one(3R,4R)-4-(Isobutoxy)-3-(triphenylmethylamino)azetidin-2-one(3S,4S)-4-(Isobutoxy)-3-(triphenylmethylamino)azetidin-2-one(3R,4R)-4-(Isobutylthio)-3-(triphenylmethylamino)azetidin-2-one(3R,4R)-4-(Isobutylsulfonyl)-3-(triphenylmethylamino)azetidin-2-one(3S,4R)-4-(Isobutoxy)-3-(triphenylmethylamino)azetidin-2-one(3S,4R)-4-(Propoxy)-3-(triphenylmethylamino)azetidin-2-one(3S,4S)-4-(Propoxy)-3-(triphenylmethylamino)azetidin-2-one(3S,4R)-4-(Benzyloxy)-3-(triphenylmethylamino)azetidin-2-one(3S,4S)-(Benzyloxy)-3-(triphenylmethylamino)azetidin-2-one(3S,4R)-4-Methoxy-3-(triphenylmethylamino)azetidin-2-one(3S,4R)-4-(Isobutenyloxy)-3-(triphenylmethylamino)azetidin-2-one(3S,4R)-4-Octyloxy-3-(triphenylmethylamino)azetidin-2-one(3S,4R)-4-Phenoxy-3-(triphenylmethylamino)azetidin-2-one(3S,4S)-4-Phenoxy-3-(triphenylmethylamino)azetidin-2-one(3S,4R)-3-[[(4-Iodophenyl)diphenylmethyl]amino]-4-(isobutoxy)azetidin-2-one(3S,4S)-4-[3-(Methoxycarbonyl)propoxy]-3-(triphenylmethylamino)azetidin-2-one(3S,4R)-4-[[2-(3-Pyridylmethyl)-1,3-dithian-2-yl]methoxy]-3-(triphenylmethylamino)-azetidin-2-one(3S,4S)-4-(Prop-2-ynyloxy)-3-(triphenylmethylamino)azetidin-2-one Methyl4-[(3S,4S)-2-oxo-3-(triphenylmethylamino)azetidin-4-yloxy]but-2-ynoateMethyl4-[(3S,4R)-2-oxo-3-(triphenylmethylamino)azetidin-4-yloxy]but-2-ynoate(3S,4R)-4-[(2(5H)Furanon-4-yl)methoxy]-3-(triphenylmethylamino)azetidin-2-one(S)-3-(Triphenylmethylamino)azetidin-2-one(RS)-3-(Triphenylmethylamino)azetidin-2-one; or(3R,4R)-4-(Methylsulfonyl)-3-(triphenylmethylamino)azetidin-2-one.
 8. Amethod for treating an inflammatory disease or disorder in a mammal inneed thereof which method comprises administering to said mammal aneffective amount of a compound of Formula

wherein Y is NH; X is O or S(O)m; m is 0 or an integer having a value of1, or 2; R₃ is optionally substituted triphenylmethyl; R₄ is optionallysubstituted C₁₋₁₀ alkyl, (CR₁₀R₂₀)_(n)C(R₁₀)=C(R₇)₂, or(CR₁₀R₂₀)n—C≡C—R₅; n is an integer having a value of 1 to 4; R₁₀ and R₂₀are independently hydrogen or C₁₋₄ alkyl; R₅ is hydrogen, alkyl, aryl,arylalkyl, heteroaryl, heteroarylalkyl, C(O)₂R₆, or C(O)R₆ wherein thealkyl, aryl, arylalkyl, heteroaryl and heteroarylalkyl moieties may beoptionally substituted; R₆ is C₁₋₁₀ alkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, heterocyclic, or heterocyclicalkyl, all of which may beoptionally substituted; R₇ is independently hydrogen, C₁₋₁₀ alkyl, aryl,arylalkyl, heteroaryl, heteroarylalkyl, heterocyclic, orheterocyclicalkyl, all of which may be optionally substituted; or apharmaceutically acceptable salt thereof.
 9. A compound of the formula:

wherein Y is NH; X is O or S R₃ is optionally substitutedtriphenylmethyl; R₄ is optionally substituted C₁₋₁₀ alkyl,(CR₁₀R₂₀)_(n)C(R_()=C(R) ₇)₂, or (CR₁₀R₂₀)n—C≡C—R₅; n is an integerhaving a value of 1 to 4; R₁₀ and R₂₀ are independently hydrogen or C₁₋₄alkyl; R₅ is hydrogen, alkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, C(O)₂R₆, or C(O)R₆ wherein the alkyl, aryl, arylalkyl,heteroaryl, and heteroarylalkyl moieties may be optionally substituted;R₆ is C₁₋₁₀ alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,heterocyclic, or heterocyclicalkyl, all of which may be optionallysubstituted; R₇ is independently hydrogen, C₁₋₁₀ alkyl, aryl, arylalkyl,heteroaryl, heteroarylalkyl, heterocyclic, or heterocyclicalkyl, all ofwhich may be optionally substituted; excluding4-methoxy-3-(triphenylmethylamino)azetidin-2-one,4-(isobutenyloxy)-3-(triphenylmethylamino)azetidin-2-one,4-(methylsulfonyl)-3-(triphenylmethylamino)azetidin-2-one, and4-(prop-2-ynyloxy)-3-(triphenylmethylamino)azetidin-2-one; or apharmaceutically acceptable salt thereof.
 10. The compound according toclaim 9 wherein R₄ is an optionally substituted C₁₋₁₀ alkyl or alkenyl.11. The compound according to claim 10 wherein the alkyl is substitutedby a ketal or thioketal.
 12. The compound according to claim 9 whereinR₄ is (CR₁₀R₂₀)n—C≡C—R₅; and R₅ is hydrogen, C(O)₂R₆ or a heteroarylring.
 13. The compound according to claim 12 wherein the heteroaryl ringis a pyridyl ring.
 14. A pharmaceutical composition comprising apharmaceutically acceptable diluent or carrier and a compound accordingto claim
 9. 15. A pharmaceutical composition comprising apharmaceutically acceptable diluent or carrier and(3S,4R)-4-(isobutenyloxy)-3-(triphenylmethylamino)azetidin-2-one, or apharmaceutically acceptable salt thereof.